apoB CRISPR Activation Plasmid (h): sc-400705-ACT

Human APOB encodes apolipoprotein B (apoB), an essential structural component of atherogenic lipoproteins that scaffolds very-low-density lipoprotein (VLDL) assembly in hepatocytes and supports conversion to low-density lipoprotein (LDL) in circulation. ApoB participates in lipid transport and cholesterol homeostasis through pathways governing lipoprotein biogenesis, secretion, and receptor-mediated uptake, integrating with endoplasmic reticulum processing and triglyceride-rich particle metabolism. Dysregulated APOB expression or apoB-containing particle production is strongly linked to hyperlipidemic states and cardiovascular disease biology, and it is also relevant to hepatic lipid handling and steatosis-associated mechanisms. As a result, APOB is widely used to model lipoprotein metabolism, apoB particle dynamics, and downstream inflammatory and vascular responses in research systems.

apoB CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous APOB expression without altering the underlying DNA sequence.

apoB CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the APOB locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the APOB transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous apoB expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native APOB locus and enabling the study of apoB-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of apoB pathway restoration in tumor cells with silenced or reduced APOB expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.